A photobleaching energy transfer analysis of CD8/MHC-I and LFA-1/ICAM-1 interactions in CTL-target cell conjugates.

Abstract:

:The photobleaching energy transfer (pbFRET) technique is a fluorescence method to measure proximity relationships between molecules, especially cell surface proteins, labeled with fluorophore-conjugated monoclonal antibodies, on a pixel-by-pixel base using digital imaging microscopy. This technique enables analysis of inter- and intramolecular proximities at cell surfaces at physiological conditions. We have developed a pbFRET approach to measure intercellular proximities in order to access spatial organization of interacting proteins in the contact region of two 'communicating' cells. Two examples, as possible application areas of this approach, are presented here: interaction between CD8 and MHC-I molecules in point contacts and interaction between LFA-1 and ICAM-1 molecules in focal contacts of CTL-target conjugates. The geometry of these protein contacts based on our resonance energy transfer (RET) data is consistent with the observed blocking effects of monoclonal antibodies (directed against the interacting proteins) on the cytolytic activity of CTLs and suggest a critical role for CD8beta-subunit in signal transmission in peripheral T-lymphocytes.

journal_name

Immunol Lett

journal_title

Immunology letters

authors

Bacsó Z,Bene L,Bodnár A,Matkó J,Damjanovich S

doi

10.1016/s0165-2478(96)02665-x

subject

Has Abstract

pub_date

1996-12-01 00:00:00

pages

151-6

issue

2-3

eissn

0165-2478

issn

1879-0542

pii

S0165-2478(96)02665-X

journal_volume

54

pub_type

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